The
mitomycin derivative 10-decarbamoyl
mitomycin C (DMC) more rapidly activates a p53-independent cell death pathway than
mitomycin C (MC). We recently documented that an increased proportion of mitosene1-beta-adduct formation occurs in human cells treated with DMC in comparison to those treated with MC. Here, we compare the cellular and molecular response of human
cancer cells treated with MC and DMC. We find the increase in
mitosene 1-beta-adduct formation correlates with a condensed nuclear morphology and increased cytotoxicity in human
cancer cells with or without p53. DMC caused more DNA damage than MC in the nuclear and mitochondrial genomes. Checkpoint 1
protein (Chk1) was depleted following DMC, and the depletion of Chk1 by DMC was achieved through the
ubiquitin proteasome pathway since chemical inhibition of the
proteasome protected against Chk1 depletion. Gene silencing of Chk1 by
siRNA increased the cytotoxicity of MC. DMC treatment caused a decrease in the level of total
ubiquitinated proteins without increasing
proteasome activity, suggesting that DMC mediated
DNA adducts facilitate signal transduction to a pathway targeting cellular
proteins for proteolysis. Thus, the mitosene-1-beta stereoisomeric
DNA adducts produced by the DMC signal for a p53-independent mode of cell death correlated with reduced nuclear size, persistent DNA damage, increased
ubiquitin proteolysis and reduced Chk1
protein.